Zhang Yejia, Phillips Frank M, Thonar Eugene J-M A, Oegema Theodore, An Howard S, Roman-Blas Jorge A, He Tong-Chuan, Anderson D Greg
Departments of Rehabilitation Medicine, Thomas Jefferson University, Philadelphia, PA 19107, USA.
Spine (Phila Pa 1976). 2008 Apr 15;33(8):831-8. doi: 10.1097/BRS.0b013e31816b1f38.
Rabbit knee articular chondrocytes overexpressing human growth factors were injected into cultured intervertebral disc explants. Survival of the injected cells and accumulation of extracellular matrix were assessed.
To define the utility of cell-based gene delivery approach for repair of the intervertebral disc.
Back pain associated with symptomatic disc degeneration is a common clinical condition. Growth factors stimulate disc cell metabolism, but the ideal method for in vivo delivery has not been established. Cells as a vehicle for delivering growth factors to the disc offer potential advantages, including prolonged production of the growth factor within the disc and vital cells to participate in the repair process.
New Zealand white rabbit articular chondrocytes transduced with adenovirus expressing human bone morphogenetic protein-7 and green fluorescence protein (GFP) (AdhBMP-7), human bone morphogenetic protein-10 and GFP (AdBMP-10), or GFP alone (AdGFP, as a control) were injected into whole disc explants. Discs were maintained in culture for 1 to 2 months. At the conclusion of the culture periods, cell survival was assessed by fluorescence microscopy and extracellular matrix accumulation was assessed with biochemical methods.
Chondrocytes achieved long-term survival in the cultured disc explants. The discs treated with chondrocytes/BMP-7 demonstrated a 50% increase in proteoglycan content within the nucleus pulposus compared to control (chondrocytes/GFP), while discs injected with chondrocytes/BMP-10 failed to show a significant increase in proteoglycan accumulation.
Our study demonstrates the ability of transduced articular chondrocytes to survive and promote proteoglycan accumulation when transplanted into the intervertebral disc. These data support the potential of a cell-based gene therapy approach for disc repair. Further studies using this approach in animal models are indicated as a step towards achieving disc repair in humans.
将过表达人类生长因子的兔膝关节软骨细胞注射到培养的椎间盘外植体中。评估注射细胞的存活情况以及细胞外基质的积累情况。
确定基于细胞的基因递送方法在椎间盘修复中的效用。
与有症状的椎间盘退变相关的背痛是一种常见的临床病症。生长因子可刺激椎间盘细胞代谢，但尚未确立理想的体内递送方法。细胞作为向椎间盘递送生长因子的载体具有潜在优势，包括在椎间盘内延长生长因子的产生以及有活力的细胞参与修复过程。
将用表达人类骨形态发生蛋白-7和绿色荧光蛋白（GFP）的腺病毒（AdhBMP-7）、人类骨形态发生蛋白-10和GFP（AdBMP-10）或仅GFP（AdGFP，作为对照）转导的新西兰白兔关节软骨细胞注射到整个椎间盘外植体中。椎间盘在培养中维持1至2个月。在培养期结束时，通过荧光显微镜评估细胞存活情况，并用生化方法评估细胞外基质积累情况。
软骨细胞在培养的椎间盘外植体中实现了长期存活。与对照组（软骨细胞/GFP）相比，用软骨细胞/BMP-7处理的椎间盘髓核中的蛋白聚糖含量增加了50%，而注射软骨细胞/BMP-10的椎间盘蛋白聚糖积累未显示出显著增加。
我们的研究证明了转导的关节软骨细胞移植到椎间盘后能够存活并促进蛋白聚糖积累。这些数据支持基于细胞的基因治疗方法在椎间盘修复方面的潜力。表明进一步在动物模型中使用这种方法作为朝着实现人类椎间盘修复迈出的一步。
